Differential coating of human red blood cells with C4 or C3 in a low ionic strength medium.
An EDTA-containing, low ionic strength medium, pH 5.1, can be used for incubation of blood to obtain coating of red blood cells with C4 or C3. The coating achieved depends on incubation temperature (4 or 37 degrees C) and on whether or not CaCl2 or MgCl2 is added to the medium. The optimal concentration of EDTA appears to be 2 mM under the present conditions. This is equimolar to the final concentration of the added CaCl/MgCl2. The choice between CaCl and MgCl2 depends on whether blood without anticoagulant or ACD blood is employed. Coated cells keep satisfactorily for 5 weeks at 4 degrees C in Alsever's solution as well as after freezing and thawing. C3-coated cells can be converted to C3d-coated ones by incubation in normal, compatible, EDTA-containing serum at 37 degrees C.
D002122:Calcium Chloride / Q000494:pharmacology; D002951:Citrates / Q000494:pharmacology; D003176:Complement C3 / Q000378:metabolism*; D003181:Complement C4 / Q000378:metabolism*; D003470:Culture Media / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D004492:Edetic Acid / Q000494:pharmacology; D004910:Erythrocyte Membrane / Q000378:metabolism*; D004912:Erythrocytes / Q000378:metabolism*; D006801:Humans; D006863:Hydrogen-Ion Concentration; D066298:In Vitro Techniques; D008274:Magnesium / Q000494:pharmacology; D009994:Osmolar Concentration; D013696:Temperature
D002951:Citrates; D003176:Complement C3; D003181:Complement C4; D003470:Culture Media; D004492:Edetic Acid; D008274:Magnesium; D002122:Calcium Chloride